LOCUS       sequence-444666-        4924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 AAV virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  AUTHORS   Kussick E, Johansen N, Taskin N, Chowdhury A, Quinlan MA, Fraser A, 
            Clark AG, Wynalda B, Martinez R, Groce EL, Reding M, Liang E, Shulga
            L, Huang C, Casper T, Clark M, Ho W, Gao Y, van Velthoven CTJ, 
            Sobieski C, Ferrer R, Berg MR, Curtis BC, English C, Day JC, Fortuna
            MG, Donadio N, Newman D, Yao S, Chakka AB, Goldy J, Torkelson A, 
            Guzman JB, Chakrabarty R, Nguy B, Guilford N, Pham TH, Wright V, 
            Ronellenfitch K, Naidoo R, Kenney J, Williford A, Ramakrishnan C, 
            Drinnenberg A, Gudsnuk K, Thyagarajan B, Smith KA, Dee N, Deisseroth
            K, Zeng H, Yao Z, Tasic B, Levi BP, Hodge R, Bakken TE, Lein ES, 
            Ting JT, Daigle TL
  TITLE     Enhancer AAVs for targeting spinal motor neurons and descending 
            motor pathways in rodents and macaque.
  JOURNAL   Cell Rep. 2025 Jun 24;44(6):115730. doi: 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21.
  PUBMED    40403722
REFERENCE   2  (bases 1 to 4924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2025-06-24"; volume: "44(6):115730. doi"; pages: " 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21"
COMMENT     Sequence Label: HCT133_rAAV-eHGT_1500m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..4924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      936..945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(942..944,945..947,948..1658)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(987..1008)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1248..1267)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1595..1616
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1751..1771)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1958..2069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2187..2327
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2187..2316
                     /label=AAV2 ITR
     rep_origin      2402..2857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2489..2508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2699..2720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2874..2893)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2993..3015
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3053..3071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3139..3243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3244..3312,3313..4104)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3462..3481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4275..4863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4764..4783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtcct gattgtcctg tgtccaggtg ttatgtcaca
      181 gaaattgaaa gactttgggg agcaagtttc ctttatgttc tttgtttcaa gtactcttta
      241 cttgtttcca tgacagagtt tttatgaaga ttaaaataag tgaaataata ccttctattt
      301 ttgtagacta atacttgata cagtgaaaaa tttcagtcaa tattaactgt tattttcaca
      361 acagaaaaaa atgactcatt gttcagtttg gcataatgaa aaatagttca agacacatat
      421 tcattctctc ctaatttcat tcctttaaaa taagggcaat acattggcat cagatgaaca
      481 gttggcttag aagagttccc aatatagact tggacaatct ccctttggac ctcctgtgtc
      541 cctctctgtc tacccaggcc ttcctcctct agctgagcat tgaaacacct gcctgcttgg
      601 caagttgaag aaggtccaat ctgctaaaat aattgaatgc ttcagaattt gccaacagta
      661 aagacagatg ctgtaaaggc tgagtgagaa atatgatctc tctttggatg cattttgttt
      721 tatctttttc cttattaaaa aaaggtgatg tgtgtaaccc cagggcctat ggaattatca
      781 tttgcccctt acctccaaca aattaaatct atattttaca ctgaaatgaa tggttatatt
      841 gcgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      901 gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg
      961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     1081 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     1141 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1381 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1441 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
     1561 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1621 ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
     1681 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1741 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1801 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     1861 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     1921 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     1981 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2041 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2101 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     2161 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     2221 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2281 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     2341 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     2401 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2461 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2521 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2581 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2641 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2701 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     2761 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     2821 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     2881 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2941 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3001 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3061 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3241 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3301 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3361 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3421 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3481 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3541 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3601 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3661 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3721 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3781 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3841 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3901 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3961 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4021 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4081 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4141 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4201 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4261 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4321 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4381 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4441 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4501 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4561 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4621 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4681 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4741 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4801 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4861 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4921 atgt
//
